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. 2013 Dec 23;19(1):23–36. doi: 10.2478/s11658-013-0112-1

CD44 and CD24 cannot act as cancer stem cell markers in human lung adenocarcinoma cell line A549

Raheleh Roudi 1,2,4, Zahra Madjd 1,3,4,✉, Marzieh Ebrahimi 2,✉, Fazel Sahraneshin Samani 2, Ali Samadikuchaksaraei 4,5
PMCID: PMC6275711  PMID: 24363164

Abstract

Cancer stem cells (CSCs) are subpopulations of tumor cells that are responsible for tumor initiation, maintenance and metastasis. Recent studies suggested that lung cancer arises from CSCs. In this study, the expression of potential CSC markers in cell line A549 was evaluated. We applied flow cytometry to assess the expression of putative stem cell markers, including aldehyde dehydrogenase 1 (ALDH1), CD24, CD44, CD133 and ABCG2. Cells were then sorted according to the expression of CD44 and CD24 markers by fluorescence-activated cell sorting (FACS) Aria II and characterized using their clonogenic and sphere-forming capacity. A549 cells expressed the CSC markers CD44 and CD24 at 68.16% and 54.46%, respectively. The expression of the putative CSC marker ALDH1 was 4.20%, whereas the expression of ABCG2 and CD133 was 0.93%. Double-positive CD44/133 populations were rare. CD44+/24+ and CD44+/CD24−/low subpopulations respectively exhibited 64% and 27.92% expression. The colony-forming potentials in the CD44+/CD24+ and CD44+/CD24−/low subpopulations were 84.37 ± 2.86% and 90 ± 3.06%, respectively, while the parental A549 cells yielded 56.65 ± 2.33% using the colony-formation assay. Both isolated subpopulations formed spheres in serumfree medium supplemented with basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). CD44 and CD24 cannot be considered potential markers for isolating lung CSCs in cell line A549, but further investigation using in vivo assays is required.

Key words: Cancer stem cells, Lung cancer, Cell line A549, Colony-formation assay, Sphere-formation assay, CD44, CD24, CD133, ALDH1, ABCG-2

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Abbreviations used

ABC

ATP-binding cassette

ADC

adenocarcinoma

ALDH1

aldehyde dehydrogenase 1

bFGF

basic fibroblast growth factor

CFU

colony-forming unit

CSCs

cancer stem cells

DEAB

diethylaminobenzaldehyde

EGF

epidermal growth factor

GFs

growth factors

LCC

large-cell carcinoma

NSCLC

non-small cell lung carcinoma

SCC

squamous-cell carcinoma

SP

side population

Contributor Information

Zahra Madjd, Phone: +982188622608, FAX: +982188622608, Email: Zahra.madjd@yahoo.com.

Marzieh Ebrahimi, Phone: +982122306485, FAX: +982122413720, Email: Mebrahimi@royaninstitute.org.

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